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© The Rockefeller University Press, 0021-9525/1998//1239 $5.00
The Journal of Cell Biology, Volume 143, Number 5, , 1998 1239-1247

Article

The "8-kD" Cytoplasmic Dynein Light Chain Is Required for Nuclear Migration and for Dynein Heavy Chain Localization in Aspergillus nidulans



Susan M. Beckwith, Christian H. Roghi, Bo Liu, and N. Ronald Morris

Department of Pharmacology, University of Medicine and Dentistry of New Jersey—Robert Wood Johnson Medical School, Piscataway, New Jersey 08854

The heavy chain of cytoplasmic dynein is required for nuclear migration in Aspergillus nidulans and other fungi. Here we report on a new gene required for nuclear migration, nudG, which encodes a homologue of the "8-kD" cytoplasmic dynein light chain (CDLC). We demonstrate that the temperature sensitive nudG8 mutation inhibits nuclear migration and growth at restrictive temperature. This mutation also inhibits asexual and sexual sporulation, decreases the intracellular concentration of the nudG CDLC protein and causes the cytoplasmic dynein heavy chain to be absent from the mycelial tip, where it is normally located in wild-type mycelia. Coimmunoprecipitation experiments with antibodies against the cytoplasmic dynein heavy chain (CDHC) and the nudG CDLC demonstrated that some fraction of the cytoplasmic dynein light chain is in a protein complex with the CDHC. Sucrose gradient sedimentation analysis, however, showed that not all of the NUDG protein is complexed with the heavy chain. A double mutant carrying a cytoplasmic dynein heavy chain deletion plus a temperature-sensitive nudG mutation grew no more slowly at restrictive temperature than a strain with only the CDHC deletion. This result demonstrates that the effect of the nudG mutation on nuclear migration and growth is mediated through an interaction with the CDHC rather than with some other molecule (e.g., myosin-V) with which the 8-kD CDLC might theoretically interact.

Key Words: cytoplasmic dynein • light chain • nucleus • migration • Aspergillus

Abbreviations used in this paper: CDHC, cytoplasmic dynein heavy chain; CDLC, cytoplasmic dynein light chain; DAPI, 4',6-diamidino-2-phenylindone; MT, microtubule; SPB, spindle pole bodies; ts, temperature sensitive.

Dr. Beckwith's present address is U.S.U.H.S., 15104 Dufief Dr., N. Potomac, MD 20878. Dr. Roghi's present address is University of Manchester, School of Biological Sciences, 205 Stopford Building, Manchester, M13 9PT, UK, and Dr. Liu's present address is Section of Plant Biology, University of California at Davis, Davis, CA 95616.

Address all correspondence to N.R. Morris, Department of Pharmacology, 675 Hoes Lane, Piscataway, NJ 08854. Tel.: (732) 235-4166. Fax: (732) 235-4073. E-mail: morrisnr{at}umdnj.edu


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